Elizabeth R. Smith, Ph.D.
Research Assistant Professor
Department of Cell Biology
University of Miami Miller School of Medicine
Miami, FL 33136
March 31, 2015
SEEDS You Choose Final Report—Training in Ovarian Granulosa Cell Biology
Summary:
The SEEDS You Choose award sponsored a visit to the laboratory of Dr. Barbara Vanderhyden at the University of Ottowa on November 28-December 3, 2014. The objective of the visit was to obtain basic training in ovarian granulosa cell biology. Our research investigates the hypothesis is that follicles, likely the granulosa cells within the follicles, produce a factor that regulates epithelial cell proliferation; depletion of follicles as found in menopause removes the factor and permits epithelial growth. I reasoned that specific training in granulosa cell biology should better allow us to approach the questionsrelated to how ovarian epithelial cells are regulated.
The visit had two parts: the first part consisted of hands on training in collecting and culturing murine ovarian granulosa cells; the second part involved extensive discussion about the purpose, applications, and directions of our individual research programs.
To demonstrate how to maximize the number and quality of granulosa cells, Dr. Vanderhyden and her technician had prepped two sets of mice before my arrival at the laboratory. One set had been stimulated with DES to increase the number of granulosacells associated with pre-antral follicles, and the second set had been injected with PMSG/hCG to increase the number of antral follicles and associated granulosa cells. DES priming yields fairly pure populations of pre-antralgranulosa cells. PMSG/hCG yields a mixture of granulosa cells, both mural and cumulus, and luteinized cells of the corpora lutea. Dr. Vanderhyden worked along side me to show me how to best isolate and propagate the cells in culture.
On the second day of the visit, I attended her lab’s weekly research meeting. She has 8 graduate students and three senior research scientists who have been in the lab for 10 years or more. Each student gave a brief presentation of his or her research, and I gave a longer one on my own studies investigating the relationship between menopausal status and ovarian cancer risk. Questions and discussion followed, and I spoke with a couple of graduate students one-on-one about similar research interests.
Outcomes:
The SEEDS award has allowed me to establish a new research direction, provided impetus for grant submissions, and fostered collaborations between the two labs.
We have since successfully established in our lab the protocols to isolate murine granulosa cells and co-culture with ovarian surface epithelial cells. The results have been fairly promising so far, in that granulosa cell conditioned medium does seem to inhibit ovarian surface epithelial cell growth. Results also suggest that the effect may be non-cell autonomous but rather requires interaction between different cell types of the ovary. We plan to continue this research direction.
I submitted in February 2015 an R21 application to the NIH-NCI entitled “Ovarian Aging and Cancer Risk”, which incorporates experiments to examine the epithelial-granulosa interactions and to identify the putative factor that may be involved in epithelial regulation.
Finally, I believe that the SEEDS sponsored visit has, in fact, fostered collaboration between Dr. Vanderhyden’s and our lab. Recently, Dr. Vanderhyden and one of her graduate students contacted us about our work investigating Disabled-2 (Dab2) in ovarian cancer. We are providing mouse ovarian surface epithelial and oviduct cells isolated from Dab2 conditional and straight knockout mice. I hope that we can continue to collaborate and stimulate productive investigations in ovarian cancer biology.